Yang Wenzhi, Zhang Jingxian, Yao Changliang, Qiu Shi, Chen Ming, Pan Huiqin, Shi Xiaojian, Wu Wanying, Guo Dean
Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, People's Republic of China.
Guangxi Wuzhou Pharmaceutical (Group) Co., Ltd., Wuzhou 543000, People's Republic of China.
J Pharm Biomed Anal. 2016 Sep 5;128:322-332. doi: 10.1016/j.jpba.2016.05.035. Epub 2016 Jun 3.
Xueshuantong Injection (XSTI), derived from Notoginseng total saponins, is a popular traditional Chinese medicine injection for the treatment of thrombus-resultant diseases. Current knowledge on its therapeutic basis is limited to five major saponins, whereas those minor ones are rarely investigated. We herein develop an offline two-dimensional liquid chromatography/quadrupole time-of-flight mass spectrometry-fast data directed analysis (offline 2D LC/QTOF-Fast DDA) approach to systematically characterize the saponins contained in XSTI. Key parameters affecting chromatographic separation in 2D LC (including stationary phase, mobile phase, column temperature, and gradient elution program) and the detection by QTOF MS (involving spray voltage, cone voltage, and ramp collision energy) were optimized in sequence. The configured offline 2D LC system showed an orthogonality of 0.84 and a theoretical peak capacity of 8976. Total saponins in XSTI were fractionated into eleven samples by the first-dimensional hydrophilic interaction chromatography, which were further analyzed by reversed-phase UHPLC/QTOF-Fast DDA in negative ion mode. The fragmentation features evidenced from 36 saponin reference standards, high-accuracy MS and Fast-DDA-MS(2) data, elemental composition (C<80, H<120, O<50), double-bond equivalent (DBE 5-15), and searching an in-house library of Panax notoginseng, were simultaneously utilized for structural elucidation. Ultimately, 148 saponins were separated and characterized, and 80 have not been isolated from P. notoginseng. An in-depth depiction of the chemical composition of XSTI was achieved. The results obtained would benefit better understanding of the therapeutic basis and significant promotion on the quality standard of XSTI as well as other homologous products.
血栓通注射液（XSTI）源自三七总皂苷，是一种用于治疗血栓性疾病的常用中药注射液。目前对其治疗基础的认识仅限于五种主要皂苷，而对那些次要皂苷的研究较少。我们在此开发了一种离线二维液相色谱/四极杆飞行时间质谱-快速数据导向分析（离线二维液相色谱/四极杆飞行时间质谱-快速数据依赖性采集）方法，以系统地表征血栓通注射液中所含的皂苷。依次优化了二维液相色谱中影响色谱分离的关键参数（包括固定相、流动相、柱温以及梯度洗脱程序）和四极杆飞行时间质谱的检测参数（包括喷雾电压、锥孔电压和斜坡碰撞能量）。所配置的离线二维液相色谱系统显示出0.84的正交性和8976的理论峰容量。血栓通注射液中的总皂苷通过一维亲水相互作用色谱法分离成11个样品，然后在负离子模式下通过反相超高效液相色谱/四极杆飞行时间质谱-快速数据依赖性采集进一步分析。从36种皂苷参考标准品、高精度质谱和快速数据依赖性采集质谱/质谱数据、元素组成（C<80，H<120，O<50）、双键当量（DBE 5-15）以及搜索三七内部库中获得的裂解特征，同时用于结构解析。最终，分离并鉴定了148种皂苷，其中80种尚未从三七中分离得到。实现了对血栓通注射液化学成分的深入描述。所得结果将有助于更好地理解其治疗基础，并显著推动血栓通注射液以及其他同类产品质量标准的提升。
